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HYB: Ploidy testing

  • Subject: HYB: Ploidy testing
  • From: vince lewonski <vincelewonski@yahoo.com>
  • Date: Fri, 4 Jan 2002 13:49:49 -0800 (PST)


 
  Eloy Boon <eloyboon@iribov.nl> wrote: 
I use flow cytometry to analyse the DNA content of Irisses. In the breesing
program I am working I convert diploid Irisses in vitro to tetraploid. In a
very early stage we take a small part of tissue and isolate nuclei and stain
them with a DNA dye, and analyse by flowcytometry the ploidy level. I
multiply the tetraploids and analyse this material again. In this way we can
exclude the mixoploid clones after two multiplication cycles.
In most crops I prefer to work with colchicine since this tends to give more
reproducable results in our lab compared to Oryzaline.


I 'am working in breeding of Dutch Iris. The most important cultivars are
crosses between I. xiphium and I. tingitana. The hybrids are sterile but
after converting them to tetraploid fertility is restored. I converted both
parent species and hybrids. During analyses we found that the DNA  content
of the diploid parents is different. The hybrid has intermediate DNA
content. If I receive unknown material I can tell on the basis of DNA
content what is the constitution of the material: A triploid with 2 sets of
tingitana has a different DNA content as an triploid with 1 set of
tingitana. I use the system also to detect in an early stage if a plant is a
hybrid after embryo culture.

It would be interesting to see if this system can also be used for other
Iris groups. 

Best regards,


Eloy Boon

Iribov
The Netherlands


V. sez:

I'll say it would be interestng!!! Few questions for Eloy:

What is the method used by you to convert using the colchicine? It sounds like your death rate isn't nearly as high as others have discussed. Or is there something different about the Dutch irises that makes conversion attempts less likely to be fatal?

Are your crosses mostly between different species (I. xiphium and I. tingitana) of Dutch irises, then? Or are named hybrids also used?








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Vince Lewonski
vincelewonski@yahoo.com
Secane, Pennsylvania, USA Zone 6b
"Hard work might not kill me, but it would sure hurt like hell!!!"


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